Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain.
A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-alpha-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue-Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disfulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected with HIV-1.
Ghetie
V
V
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
Till
M A
MA
Ghetie
M A
MA
Uhr
J W
JW
Vitetta
E S
ES
D015704:CD4 Antigens / Q000008:administration & dosage*; D002853:Chromatography, Liquid; D006801:Humans; D000922:Immunotoxins / Q000302:isolation & purification* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000008:administration & dosage; D012276:Ricin / Q000008:administration & dosage*
D015704:CD4 Antigens; D000922:Immunotoxins; D011994:Recombinant Proteins; D012276:Ricin